HardyCHROM™ UTI

Cat. no. G313 HardyCHROM™ UTI, 15x100mm Plate, 18ml 10 plates/bag

Cat. no. J119 Blood Agar / HardyCHROM™ UTI, 15x100mm Biplate, 10ml/10ml 10 plates/bag

INTENDED USE
HardyCHROM™ UTI is a non-selective chromogenic medium recommended for the cultivation, differentiation and
enumeration of urinary tract pathogens based on colony color and morphology.

SUMMARY
There are a number of organisms routinely isolated from urinary tract infections (UTI). Most UTIs are caused by
Escherichia coli alone, or in combination with other organisms. The most frequently isolated species produce
characteristic enzymes. Chromogenic substrates (chromogens) incorporated into HardyCHROM™ UTI produce
different colored compounds when they are degraded by specific microbial enzymes. Thus, HardyCHROM™ UTI can
be used for the cultivation and differentiation of various groups of organisms with only a minimum number of
confirmatory tests.

Peptones supply the necessary nutrients, and the mixture of chromogens permits detection and differentiation of the
isolated organisms. This medium contains no inhibitory substances and is not selective. The swarming of Proteus is
partially to completely inhibited.

FORMULA
Ingredients per liter of deionized water:*
Peptones 16.0gm
Chromogenic Mixture 5.0gm
Agar 15.0gm

Final pH 6.9 +/- 0.3 at 25°C.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-8°C away from direct light. Media should not be used if there are any signs of
deterioration (shrinking, cracking, or discoloration), contamination, or if the expiration date has passed.
Chromogens are especially light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.
The expiration date applies to the product in its intact packaging when stored as directed.
This product has the following shelf life from the date of manufacture:
90 Days: G313 HardyCHROM™ UTI
J119 Blood Agar / HardyCHROM™ UTI
Refer to the document "Storage" on the Hardy Diagnostics Technical Document website for more information.

PRECAUTIONS
This product may contain components of animal origin. Certified knowledge of the origin and/or sanitary state of the
animals does not guarantee the absence of transmissible pathogenic agents. Therefore, it is recommended that these
products be treated as potentially infectious and handled observing the usual universal blood precautions. Do not ingest,
inhale, or allow to come into contact with skin.

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This product is for in vitro diagnostic use only and is to be used only by adequately trained and qualified laboratory
personnel. Observe approved biohazard precautions and aseptic techniques. All laboratory specimens should be
considered infectious and handled according to "standard precautions". The "Guideline for Isolation Precautions" is
available from the Centers for Disease Control and Prevention at www.cdc.gov/ncidod/dhqp/gl_isolation.html.
For additional information regarding specific precautions for the prevention of the transmission of all infectious agents
from laboratory instruments and materials, and for recommendations for the management of exposure to infectious
disease, refer to CLSI document M-29: Protection of Laboratory Workers from Occupationally Acquired Infections:
Approved Guideline.
Sterilize all biohazard waste before disposal.

Refer to the document "Precautions When Using Media" on the Hardy Diagnostics Technical Document website for
more information.

Refer to the SDS Search instructions on the Hardy Diagnostics website for more information.

PROCEDURE
Specimen Collection: Consult listed references for information on specimen collection.(2-4) Infectious material should be
submitted directly to the laboratory without delay and protected from excessive heat and cold. If there is to be a delay in
processing, the specimen should be refrigerated until inoculation.

Consult the listed references for information regarding the processing of specimens.(1-5)

Protect media from light during storage and incubation as the product is light sensitive.

Method of Use: Allow the plates to warm to room temperature. The agar surface should be to dry prior to inoculating.
Inoculate and streak the specimen as soon as possible after collection. For quantitative testing streak plate with 0.01ml
(Cat. no. HS10R) or 0.001ml calibrated loop (Cat. no. HS1R). Incubate plates in an inverted position, aerobically at 35
+/- 2°C for no less than 24 hours. Examine plates for colonies showing typical morphology and color after 24 hours, but
no later than 36 hours.

Do not incubate in an atmosphere supplemented with CO 2 .

INTERPRETATION OF RESULTS
After incubation, the plates should show isolated colonies. Isolated colonies are necessary for demonstration of typical
color and morphology.

Escherichia coli produces medium to large sized colonies that are rose to magenta in color, with darker pink centers. No
further testing is needed.

Colonies that resemble E. coli (pink to rose), but are small or pinpoint in size, require further identification procedures
such as the Indole Spot Test (DMACA, Cat. no. Z65). See "Limitations" section below.

Enterococcus spp. appear as small, teal to turquoise colored colonies. No further testing is needed.

Proteus, Morganella, and Providencia spp. produce clear to light yellow colonies with golden-orange halo diffused
through surrounding media. Additionally, approximately 50% of Proteus vulgaris isolates will produce blue-green or
green colonies with a golden-orange halo. Proteus vulgaris can be identified by a positive spot indole test (Cat. no. Z65).
Further biochemical tests are needed for complete identification of the other members of this group. Indole Spot Test
(Cat. no. Z65) may be performed from the plate. H 2 S production and ornithine decarboxylase (Cat. no. Y44) permit
differentiation of the genera.

Klebsiella, Enterobacter, and Serratia spp. produce large, deep blue or dark indigo colonies. Citrobacter spp. produce
dark blue colonies often with a rose halo in the surrounding media. Further biochemical tests such as the Microgen GN-
A Panel (Cat. no. MID64) are needed for complete identification.

Pseudomonas spp. produce colorless to light yellow-green, translucent colonies which may have a slight iridescence with
crinkled edges. Further biochemical tests, including an oxidase test (Cat. no. Z93) may be needed for complete
identification.
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Staphylococcus aureus produce opaque, cream to white colored colonies. Note: Colonies may turn pink after 72 hours.
Further tests (StaphTEX™, Cat. no. ST50) are needed for complete identification.

Staphylococcus saprophyticus produce opaque, pink colonies. Further tests, such as novobiocin-resistance (Cat. no.
Z7291), are needed for complete identification.

Staphylococcus epidermidis grows as small, white colonies. Further biochemical tests are needed for complete
identification.

Candida albicans, Candida tropicalis, and Candida glabrata produce small, opaque, white, moist colonies. Further
biochemical tests such as AlbiQuick™ (Cat. no. Z121) or HardyCHROM™ Candida (Cat. no. G301) are needed for
complete identification.

Candida krusei appears as small, white, dry colonies which have a rough appearance. Further biochemical tests are
needed for complete identification.

Listeria monocytogenes or other Listeria spp. may be present in urine. Colonies of Listeria are very small, blue to blue-
green colonies. Perform a Gram stain of organisms producing small, blue to blue-green colonies that are PYR-negative.
The presence of gram-positive bacilli is suggestive of Listeria spp. but further biochemical tests are necessary for
complete identification.

Streptococcus agalactiae isolated from urine appears as very small clear blue colonies, very small clear white colonies or
very small pink or pink-blue colonies. Further tests, such as Strep B Carrot Broth™ (Cat. no. Z140), are needed for
complete identification.

For organisms other than E. coli and Enterococcus spp. biochemical tests should be performed on colonies from pure
cultures for complete identification. Use a filter paper to perform rapid tests. Do not apply any detection reagents directly
on the colonies growing on the medium.

Organism Description Photo Color

rose to magenta colonies

E. coli
with darker pink centers

Klebsiella, Enterobacter, and deep blue or dark indigo

Serratia spp. colonies

Enterococcus spp. teal to turquoise colonies

clear to light yellow

colonies with golden-
Proteus, Morganella, and orange halo in the
Providencia spp. surrounding media (some
Proteus vulgaris colonies
will be blue-green)

opaque, cream to white

Staphylococcus aureus
colored colonies

colorless to light yellow-

Pseudomonas spp.
green colonies

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 Staphylococcus saprophyticus opaque, pink colonies

Candida albicans, Candida krusei, small, opaque, white,

Candida tropicalis, and Candida moist colonies (C. krusei
glabrata will be a rough colony)

dark blue colonies often

Citrobacter spp. with a rose halo in the
surrounding media

LIMITATIONS
Color-blind individuals may encounter difficulty in distinguishing the color differences on HardyCHROM™ UTI.
Use of this medium for non-clinical or clinical specimens other than urine has not been validated.
Some rare strains of C. freundii may produce small, pink or rose colored colonies, with color similar to E. coli. To
prevent misidentification, a rapid Indole Spot Test (Cat. no. Z65) may be performed since C. freundii is indole-negative
and E. coli is indole-positive.
Aerococcus urinae does not grow well on this medium. After 48 hours the colonies are very small to pinpoint and are
colorless.
Corynebacterium renale does not grow on this medium (48 hours).
Do not use Kovacs Indole Reagent on dark rose or pink colonies as the colony color may interfere with the red color of a
positive indole reaction. Use only dimethylaminocinnamaldehyde (DMACA - Indole Spot Reagent, Cat. no. Z65) for
indole testing.
This medium is non-selective so most UTI pathogens will grow. Colonies that are clear and do not react with the
chromogenic substrates must be further tested with appropriate biochemical or serological tests for definitive
identification. Fastidious organisms such as Mycoplasma, Neisseria, and Haemophilus cannot grow on this medium.
Minimize exposure of HardyCHROM™ UTI medium to light before and during incubation, as light can destroy the
chromogens.
In rare cases, naturally-occurring pigmented strains may produce colony colors that may interfere with typical color
reactions. Therefore, it is recommended that biochemical, immunological, molecular, or mass spectrometry testing be
performed on these colonies from pure culture for complete identification.
Refer to the document "Limitations of Procedures and Warranty" on the Hardy Diagnostics Technical Document website
for more information.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, other culture media, swabs, applicator sticks,
incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

The following organisms are routinely used for testing at Hardy Diagnostics:

Inoculation Incubation
Test Organisms Results
Method* Time Temperature Atmosphere
Growth; medium sized
Escherichia coli rose to magenta
A 24hr 35°C Aerobic
ATCC® 25922 colonies with darker
pink centers
Growth; large, deep
Klebsiella pneumoniae
A 24hr 35°C Aerobic blue or dark indigo
ATCC® 13883
colonies

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 Enterococcus faecalis Growth; small, teal to
A 24hr 35°C Aerobic
ATCC® 29212 turquoise colonies
Growth; clear to light
yellow colonies with
Proteus mirabilis
A 24hr 35°C Aerobic golden-orange color
ATCC® 12453
diffused through
surrounding media
Growth; opaque, cream
Staphylococcus aureus
A 24hr 35°C Aerobic to white colored
ATCC® 25923
colonies
Growth; colorless to
light yellow-green,
Pseudomonas aeruginosa
A 24hr 35°C Aerobic translucent colonies,
ATCC® 27853
which may have a slight
iridescence
Staphylococcus
Growth; opaque, pink
saprophyticus A 24hr 35°C Aerobic
colonies
ATCC® 15305
Candida albicans Growth; small, white,
A 24hr 35°C Aerobic
ATCC® 10231 moist colonies
Growth; dark blue
Citrobacter freundii colonies, often with a
A 24hr 35°C Aerobic
ATCC® 8090 rose halo in the
surrounding media
* Refer to the document "Inoculation Procedures for Media QC" on the Hardy Diagnostics Technical Document website
for more information.

USER QUALITY CONTROL

Check for signs of contamination and deterioration. Users of commercially prepared culture media may be required to
perform quality control testing to demonstrate growth or a positive reaction and to demonstrate inhibition or a negative
reaction (where applicable). Refer to the following documents on the Hardy Diagnostics Technical Document website
for more information on QC: "Introduction to Quality Control," "Finished Product Quality Control Procedures," or "The
CLSI Standard and Recommendations for User Quality Control of Media." Also see the following reference for more
specific information. (1-5)

PHYSICAL APPEARANCE
HardyCHROM™ UTI should appear translucent, and light off-white in color.

Escherichia coli (ATCC® 25922) colonies growing on Klebsiella pneumoniae (ATCC® 13883) colonies
HardyCHROM™ UTI (Cat. no. G313). Incubated growing on HardyCHROM™ UTI (Cat. no. G313).
aerobically for 24 hours at 35°C. Incubated aerobically for 24 hours at 35°C.

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Enterococcus faecalis (ATCC® 29212) colonies growing Proteus mirabilis (ATCC® 12453) colonies growing
on HardyCHROM™ UTI (Cat. no. G313). Incubated on HardyCHROM™ UTI (Cat. no. G313). Incubated
aerobically for 24 hours at 35°C. aerobically for 24 hours at 35°C.

Staphylococcus aureus (ATCC® 25923) colonies Pseudomonas aeruginosa (ATCC® 27853) colonies
growing on HardyCHROM™ UTI (Cat. no. G313). growing on HardyCHROM™ UTI (Cat. no. G313).
Incubated aerobically for 24 hours at 35°C. Incubated aerobically for 24 hours at 35°C.

Staphylococcus saprophyticus (ATCC® 15305) colonies Candida albicans (ATCC® 10231) colonies growing
growing on HardyCHROM™ UTI (Cat. no. G313). on HardyCHROM™ UTI (Cat. no. G313). Incubated
Incubated aerobically for 24 hours at 35°C. aerobically for 24 hours at 35°C.

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Citrobacter freundii (ATCC® 8090) colonies growing Lactobacillus fermentum (ATCC® 9338), left, and
on HardyCHROM™ UTI (Cat. no. G313). Incubated Lactobacillus acidophilus (ATCC® 4356), right, colonies
aerobically for 24 hours at 35°C. growing on HardyCHROM™ UTI (Cat. no. G313).
Incubated aerobically for 24 hours at 35°C.

Clinical strain of group B streptococci (GBS) growing Clinical strains of group C streptococci growing on
on HardyCHROM™ UTI (Cat. no. G313). Incubated HardyCHROM™ UTI (Cat. no. G313). Incubated
aerobically for 24 hours at 35°C. aerobically for 24 hours at 35°C.

Streptococcus agalactiae (ATCC® 13813, non-

hemolytic), left, and Streptococcus agalactiae (ATCC®
12386, hemolytic), right, colonies growing on
HardyCHROM™ UTI (Cat. no. G313). Incubated
aerobically for 24 hours at 35°C.

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REFERENCES
1. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed., A.S.
Weissfeld. American Society for Microbiology, Washington, D.C.
2. Versalovic, J., et al. Manual of Clinical Microbiology. American Society for Microbiology, Washington, D.C.

3. Tille, P.M., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

4. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology,
Washington, D.C.

5. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory
Standards Institute (CLSI - formerly NCCLS), Wayne, PA.

6. Merlino, J., et al. Journal of Clinical Microbiology, American Society for Microbiology; 35:1788-1793.

ATCC is a registered trademark of the American Type Culture Collection.

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HARDY DIAGNOSTICS
1430 West McCoy Lane, Santa Maria, CA 93455, USA
Phone: (805) 346-2766 ext. 5658
Fax: (805) 346-2760
Website: www.HardyDiagnostics.com
Email: TechService@HardyDiagnostics.com

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Copyright© 1996 - 2015 by Hardy Diagnostics. All rights reserved.

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